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Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells.

Publication ,  Journal Article
Stephens, SB; Dodd, RD; Lerner, RS; Pyhtila, BM; Nicchitta, CV
Published in: Methods Mol Biol
2008

All eukaryotic cells display a dramatic partitioning of mRNAs between the cytosol and endoplasmic reticulum (ER) compartments-mRNAs encoding secretory and integral membrane proteins are highly enriched on ER-bound ribosomes and mRNAs encoding cytoplasmic/nucleoplasmic proteins are enriched on cytosolic ribosomes. In current views, this partitioning phenomenon occurs through positive selection-mRNAs encoding signal sequence-bearing proteins are directed into the signal recognition particle pathway early in translation and trafficked as mRNA/ribosome/nascent polypeptide chain complexes to the ER. In the absence of an encoded signal sequence, mRNAs undergo continued translation on cytosolic ribosomes. Recent genome-wide analyses of mRNA partitioning between the cytosol and the ER compartments have identified subsets of mRNAs that are non-canonically partitioned to the ER-although lacking an encoded signal sequence, they are translated on ER-bound ribosomes. These findings suggest that multiple, and as yet unidentified, pathways exist for directing mRNA partitioning in the cell. In this contribution, we briefly review the literature describing the subcellular partitioning patterns of mRNAs and present a detailed methodology for studying this fundamental, yet poorly understood process.

Duke Scholars

Published In

Methods Mol Biol

DOI

ISSN

1064-3745

Publication Date

2008

Volume

419

Start / End Page

197 / 214

Location

United States

Related Subject Headings

  • Ribosomes
  • RNA, Messenger
  • Oligonucleotide Probes
  • NIH 3T3 Cells
  • Mice
  • Jurkat Cells
  • Humans
  • Hela Cells
  • HeLa Cells
  • Endoplasmic Reticulum
 

Citation

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Stephens, S. B., Dodd, R. D., Lerner, R. S., Pyhtila, B. M., & Nicchitta, C. V. (2008). Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells. Methods Mol Biol, 419, 197–214. https://doi.org/10.1007/978-1-59745-033-1_14
Stephens, Samuel B., Rebecca D. Dodd, Rachel S. Lerner, Brook M. Pyhtila, and Christopher V. Nicchitta. “Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells.Methods Mol Biol 419 (2008): 197–214. https://doi.org/10.1007/978-1-59745-033-1_14.
Stephens SB, Dodd RD, Lerner RS, Pyhtila BM, Nicchitta CV. Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells. Methods Mol Biol. 2008;419:197–214.
Stephens, Samuel B., et al. “Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells.Methods Mol Biol, vol. 419, 2008, pp. 197–214. Pubmed, doi:10.1007/978-1-59745-033-1_14.
Stephens SB, Dodd RD, Lerner RS, Pyhtila BM, Nicchitta CV. Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells. Methods Mol Biol. 2008;419:197–214.

Published In

Methods Mol Biol

DOI

ISSN

1064-3745

Publication Date

2008

Volume

419

Start / End Page

197 / 214

Location

United States

Related Subject Headings

  • Ribosomes
  • RNA, Messenger
  • Oligonucleotide Probes
  • NIH 3T3 Cells
  • Mice
  • Jurkat Cells
  • Humans
  • Hela Cells
  • HeLa Cells
  • Endoplasmic Reticulum